Prostatitis syndromes cause major morbidity with a 10 percent prevalence among adult men. This project focuses on the most common category, chromc prostatitis/chronic pelvic pain syndrome (CP/CPPS). Long-term objectives are to determine the causes, consequences, and optimal therapy. Our working model is that bacterial infection is critical in many patients. Specific Aim #1. 16S rDNA evaluation, cloning, sequencing and phylogenitic studies. We will test the hypothesis that CP/CPPS patients have prostatic bacteria that distinguish them from controls. In pilot studies, patients with expressed prostatic secretion (EPS) inflammation (WBCs) were more likely to have bacterial rthosomal-encoding DNAs (16S rDNAs) than those without WBCs. We will clone and sequence 16S rDNAs from patients and controls. Sequences will be compared to available databases using BLAST searches and phylogeny software. These data will allow us to determine which bacteria are most specific to CP/CPPS, and, thus, which should be targeted in clinical trials. Specific Aim #2. Bacterial viability and clinical characteristics of CP/CPPS patients. We will test the hypothesis that bacterial viability correlates with the clinical severity of CP/CPPS. In pilot studies to evaluate bacterial viability, we developed quantitative assays for bacterial elongation messenger RNAs (tuf mRNAs) and documented that some CP/CPPS patients were tuf mRNApositive. We will compare clinical characteristics of patients with 16S rDNA and tuf mRNA, patients with 1 6S rDNAs but no tuf mRNA, and those without 1 6s rDNA or tuf mRNA. The tuf mRNA studies will be correlated with improved cultutes of the same specimens. This study will provide insights into the potential value of antimicrobial therapy and identify characteristics that distinguish patients most likely to respond. Specific Aim #3. Comparison of prostatic bacteria with EPS and seminal fluid (SFA) bacteria. We will test the hypotheses that CP/CPPS patients with prostatic bacteria have similar bacteria in their EPS and/or seminal fluid (SFA) and, further, that these bacteria differ from the bacteria in EPS and/or SFA of controls. Our preliminary studies identified the most common bacteria in CP/CPPS patients' prostatic parenchyma. To show that CP/CPPS patients have similar bacteria in their SFA and/or EPS, we will determine homology of 16S rDNAs in EPS, SFA, and prostate biopsy material from individual patients. To show that these bacteria differ from the bacteria in controls, we will compare l6S rDNAs in SFA and EPS of CP/CPPS with controls. These studies will determine if EPS or SFA can be used to identify prostatic bacteria and may result in clinical methods for non-invasive diagnosis of prostatic infection.